The need for novel, minimally invasive diagnostic, prognostic, and therapeutic biomedical devices has garnered increased interest in recent years. Microneedle (MN) technology has stood out as a promising new method for drug delivery, as well as extraction of interstitial fluid (ISF). ISF comprises a large portion of the extracellular fluid in living organisms yet remains inadequately characterized for clinical applications. Current MN research has focused on the fabrication of needles with different materials like silicone, carbon, and metals. However, little effort has been put forth into improving MN holders and patches that can be used with low cost MNs, which could effectively change how MNs are attached to the human body. Here, we describe different 3D-printed MN holders, printed using an MJP Pro 2500 3D printer, and compare the ISF extraction efficiencies in CD Hairless rats. We varied design parameters that may affect the skin-holder interface, such as throat thickness, tip curvature, and throat diameter. MN arrays, with insertion depths of 1500 lm, had extraction efficiencies of 0.44 ± 0.35, 0.85 ± 0.64, 0.32 ± 0.21, or 0.44 ± 0.46 ll/min when designed with flat, concave, convex, or bevel profile geometries, respectively. Our results suggest ISF extraction is influenced by MN holder design parameters and that a concave tip design is optimal for extracting ISF from animals. The future direction of this research aims to enable a paradigm in MN design that maximizes its efficiency and engineering performance in terms of volume, pressure, and wearability, thereby automatizing usage and reducing patient intervention to ultimately benefit remote telemedicine.